Dairy Microbiology outline

Introduction and Significance of Dairy Microbiology

• Dairy Microbiology: The study of microorganisms that are associated
with milk and milk products in all aspects is defined as Dairy
Microbiology.
• Milk hygiene: Milk hygiene is concerned with the production of clean,
wholesome milk that is free from bacteria or other disease-causing
micro-organisms and the maintenance of this condition from the farm
to the consumers.
• The nutritional qualities of milk make it a desirable food for humans
and other young animals.
• However, these nutritional values also permit the growth of many
microorganisms, some of which cause undesirable changes in milk and
its products.
• Milk’s sanitary qualities are influenced by many factors in the course
of production, processing, and delivery to the consumers .
Significance of Microbes in Milk

• Information on microbial load can be used to assess its sanitary

quality and the conditions of production.
• Bacteria, if permitted to multiply, cause spoilage of milk.
• Milk is potentially susceptible to contamination with pathogens,
however, precautions must be taken to destroy them.
• Certain microbes produce chemical changes that are desirable in the
production of cheese, yogurt, and fermented milk products.
Morphology and Classification of Dairy Bacteria
1. Classification Based on Shape, Size, and Arrangement of Cells
2. Classification Based on Temperature
3. Classification Based on Oxygen Requirement
4. Physiological Grouping:
a. Acid producers
b. Gas producers
c. Proteolytic
d. Lipolytic
e. Flavor producing
f. Sweet curdling
g. Ropiness
h. Color producing
Classification based on shape, size, and arrangement of cells

1 Coccus
Bacterial cells with spherical or ellipsoidal shapes are called cocci and may
have the following type of arrangement of cells:
a) Diplococci: Cells attached in pairs (e.g. Neisseria)
b) Streptococci: Occurs in the form of chains (e.g. Streptococcus)
c) Tetrads: Form 4 cells (e.g. Pediococci)
d) Sarcinae: Form a cuboidal arrangement
e) Staphylococci: Produces bunches of cocci; grape-like clusters.
Based on shape, size..
2. Bacilli
Bacteria with cylindrical or rod-like cells are called bacilli and these may have the
following type of arrangement of cells.
a) Diplobacilli: Cells remain attached in pairs.
b) Streptobacilli: Cells remain attached after some divisions, in form of chains.
3. Spirilla
• Bacteria with spiral or helical-shaped cells are called spirilla and may have the
following characteristic shape.
• Vibrio: rod-shaped cells having single curves
• Spirochetes: rod-shaped cells having many curves
Classification based on temperature
1 Mesophilic
Microorganisms capable of growing between 20 and 40 ℃ with an optimum growth temperature
of 37℃ are termed Mesophiles. All pathogenic organisms are mesophilic in nature
(e.g. S. aureus, E. coli).
2 Psychotropic
Microorganisms that are capable of growing at or below 7 ℃(refrigerated) but the optimum
growth temperature between 15 and 20 ℃ are termed psychotropic. These are the significant
spoilage organisms of refrigerated milk and milk products
(e.g. Pseudomonas spp., Alkaligenes spp.).
3. Thermophilic
Organisms capable of growing over 50 ℃ with an optimum growth temperature of 55 ℃ are
termed thermophiles. They are the important organisms causing outbreaks in heat-processed milk
and milk products. (e.g. Bacillus stearothermophilus, Streptococcus thermophilus)
4. Thermodurics
Organisms capable of withstanding pasteurizing temperatures of 63 ℃ /30 min. with optimum
growth temperature of 35-37℃ are termed thermoduric. They form important flora of pasteurized
or heat-processed foods (e.g. Micrococcus varians).
Classification Based on Oxygen Requirement
1 Aerobic
Organisms capable of growing in the presence of oxygen are termed aerobic organisms.
They are more efficient in the utilization of available nutrients (e.g. Bacillus spp.).
2 Anaerobic
Organisms which cannot grow in the presence of oxygen but can grow in the presence of
CO2 are termed as anaerobic organisms. They are however poisoned by O2. Some tolerate
low concentrations of O2. They produce catalase and peroxidase enzymes e.g.
Clostridium perfringens.
3 Facultative
Organisms that can grow either in the presence or absence of oxygen are termed as
facultative organisms g. E. coli, Lactococcus lactis.
4 Microaerophilic
Organisms that grow best at 1-15% of O2 levels are known as microaerophilic. They can use
O2 for energy-yielding reactions but cannot withstand more than 21% of
O2 (e.g. Campylobacter jejuni).
Physiological Grouping
1. Acid producers
The organisms capable of fermenting lactose to form lactic acid are known as acid producers. Lactic acid
coagulates milk by producing precipitation of Casein at 4.6 pH. E.g. Lactic Acid Bacteria (LAB)
i) Homofermentative LAB: Produce only lactic acid as the end product e.g. Lactococcus spp.
ii) Heterofermentative LAB: Produce equal amounts of lactic acid, carbon dioxide, acetate, and ethanol
as the major end products. e.g, few Lactobacilli, Leuconostoc spp.
2. Gas producers
Gas producers are the organisms capable of producing CO2 or \ and H2 from lactose fermentations (e.g. E.
coli, yeasts, and Clostridium spp.).
3. Proteolytic
Proteolytic organisms degrade milk proteins into soluble components by enzymes known as proteinases or
proteases (e.g. Bacillus spp, Pseudomonas spp.).

4. Lipolytic
Lipolytic organisms are capable of attacking milk fat by enzymes such as lipases liberating glycerides and
fatty acids (e.g. Pseudomonas spp, Achromobacter) Lipolytic
Molds: Geotrichum candidum, Penicillium roqueforti).
Physiological grouping….
5 Sweet curdling
Organisms capable of causing curdling of milk by rennin like enzymes before
the development of sufficient acidity are known as sweet curdling organisms
(e.g. B. subtilis, B. Cereus, and Enterococcus liquefaciens).
6 Ropiness
Ropiness organisms produce changes in the viscosity of milk or form threads
when the milk is poured from one container to another, due to the production
of gums, mucins, etc. (e.g. Alcaligenes viscosus).
7 Flavour producing
Many organisms are capable of producing different flavors in milk during
their growth. Some of the flavors and associated organisms are listed below:
• Fruity: Pseudomonas fragi
• Malty: Lactococcus lactis subsp. maltigenes
• Fishy: Proteus icthyosmius
• Unclean: E. coli
Physiological grouping
8 Colour fermentations: Many organisms are capable of producing different colors in milk
during their growth by producing chromogenic compounds. Some of the colors and
associated organisms are listed below:
• Yellow: Pseudomonas synxantha
• Blue: Pseudomonas cyanogenes
• Green: Penicillium roqueforti
• Black: Pseudomonas nigrifaciens

9. Aroma Producing: These bacteria produce desirable aroma and flavor in milk and milk
products, utilizing citric and lactic acid. Eg. Streptococcus diacetilactis, Streptococcus
citrovorus, Streptococcus parasitovorous.
Bacterial growth curve
When microorganisms are grown in a suitable liquid medium (batch culture or closed system)
and incubated growth follows a definite process. If bacterial counts are carried out at intervals
after inoculation and plotted in relation to time, a growth curve is obtained.
The typical growth curve is divided into the following phase:
1. Lag phase
2. Log phase or exponential phase
3. Stationary phase
4. Death or decline phase

Generation time: Generation time is the time required for bacteria to grow and divide i.e. one
complete cell division. Some microbes are able to divide as rapidly (by binary fission) as once
every 12 to 15 minutes, others require up to several hours, and a few very slow-growing
bacteria may require more than 24 hours per cell division. Bacterial populations are expressed
as colony-forming units per ml (CFU/ml).
 Bacterial growth curve..
1. Lag phase: During the lag phase, bacteria take some time to adapt themselves to
the new growth conditions. The lag phase is characterized by:
• No cell division
• No increase in the number of cells.
• Increase in size of bacteria
• Synthesis of RNA, enzymes, and co-enzymes for physiological activities.
2. Log phase/Exponential: Microorganisms start dividing at a constant rate.
• Bacterial cell numbers double with time
• Rate of growth remains constant
• Bacteria have the smallest size
3. Stationary phase: Cell division stops due to nutrient exhaustion and accumulation
of toxic products. The viable count remains stationary as equilibrium exists between
the dying cells and the newly formed cells. Production of antibiotics such as
Penicillin, streptomycin, etc, and enzymes by certain bacteria occur during this
phase. In endospore-forming bacteria, sporulation occurs as the bacteria enter the
stationary phase.
Bacterial growth curve…
• Death phase: This is the phase when the population decreased due to
cell death. Since it is a closed system, there is no way to add nutrients
or remove waste products. Eventually, this leads to unfavorable
conditions and a decrease in the number of living cells in the
population.
 Yeasts
• Yeasts are eukaryotic, unicellular organisms. Yeast sizes vary greatly,
depending on species and environment, typically measuring 3–
4 µm in diameter, although some yeasts can grow to 40 µm in size.
• Most yeasts reproduce asexually by mitosis, and many do so by the asymmetric
division process known as budding. Yeasts may be divided into two groups
according to their method of reproduction:
Budding: called Fungi imperfecti or false yeasts
Budding and spore formation: called Ascomycetes or true yeasts
• With their single-celled growth habit, yeasts can be contrasted with molds,
which grow hyphae. Fungal species that can take both forms (depending on
temperature or other conditions) are called dimorphic fungi.
• Yeasts are very common in the environment, and are often isolated from sugar-rich materials.
• The yeast species Saccharomyces cerevisiae converts carbohydrates to carbon
dioxide and alcohols through the process of fermentation. Kefir and kumis are milk products
made by fermenting milk with yeast and bacteria.
 Molds
• Molds are multinucleated, filamentous fungi composed of hyphae. A hypha is a branching,
tubular structure from 2-10 µm in diameter and is usually divided into cell-like units by
cross-walls called septa. The total mass of hyphae is termed mycelium. The portion of the
mycelium that anchors the mold and absorbs nutrients is called the vegetative
mycelium; the portion that produces asexual reproductive spores is termed the aerial
mycelium.
• Molds possess a rigid polysaccharide cell wall composed mostly of chitin and, like all fungi,
are eukaryotic. Molds reproduce primarily by means of asexual reproductive spores such
as conidiospores, sporangiospores, and arthrospores. These spores are disseminated by air,
water, animals, or objects and upon landing in a suitable environment, germinate and
produce new hyphae. Molds may also reproduce by means of sexual spores such as
ascospores and zygospores, but this is not common.